Anti-breast cancer synthetic peptides derived from the Anabas testudineus skin mucus fractions

Abstract

Previous study has shown the antimicrobial activities of mucus protein extracted from Anabastestudineus. In this study, we are interested in characterizing the anticancer activity of the A.testudineus antimicrobial peptides (AMPs). The mucus was extracted, fractioned, and subjected to antibacterial activity testing to confirm the fish's AMPs production. The cytotoxic activity of each fraction was also identified. Fraction 2 (F2), which shows toxicity against MCF7 and MDA-MB-231 were sent for peptide sequencing to identify the bioactive peptide. The two peptides were then synthetically produced and subjected to cytotoxic assay to prove their efficacy against cancer cell lines. The IC50 for AtMP1 against MCF7 and MDA-MB-231 were 8.25 ± 0.14 μg/ml and 9.35 ± 0.25 μg/ml respectively, while for AtMP2 it is 5.89 ± 0.14 μg/ml and 6.97 ± 0.24 μg/ml respectively. AtMP1 and AtMP2 treatment for 48 h induced breast cancer cell cycle arrest and apoptosis by upregulating the p53, which lead to upregulate pro-apoptotic BAX gene and downregulate the anti-apoptotic BCL-2 gene, consequently, trigger the activation of the caspase-3. This interaction was supported by docking analysis (QuickDBD, HPEPDOCK, and ZDOCK) and immunoprecipitation. This study provided new prospects in the development of highly effective and selective cancer therapeutics based on antimicrobial peptides.

Porcupine quills keratin peptides induces G0/G1 cell cycle arrest and apoptosis via p53/p21 pathway and caspase cascade reaction in MCF‐7 breast cancer cells

“…The results of Western blotting and PCR also cross validated that PQKPs remarkably boosted the Bax and cytochrome c secretion levels and decreased the Bcl‐2 protein secretion level in MCF‐7 breast cancer cells. Additionally, it has also been shown that the peptides derived from the skin mucus of Anabas testudineus promoted apoptosis of breast cancer cells through mitochondrial pathway and caspase cascade reaction, which was consistent with the results of this study 13 . The p53, as a powerful tumor suppressor, enhances the mitochondrial membrane permeability by activating the Bax protein, resulting in the cytochrome c release and caspase cascade occurrence 48 .…”

“…found that the peptides obtained from tuna cooking juice induced the MCF‐7 cells apoptosis by enhancing the expression levels of B‐cell lymphoma (Bcl‐2) and caspase 9, and decreasing the expression levels of Bax and cleaved‐caspase 3 12 . The bioactive peptides that were extracted from the skin mucus of Anabas testudineus also increased the level of p53 expression and caused cell cycle arrest at the G0/G1 phase in breast cancer cells 13 . Two dogfish peptides that have been isolated have shown anti‐breast cancer activity by altering the cytoskeleton and causing early autophagy suppression 10 .…”

Recent advances of bioactive proteins/polypeptides in the treatment of breast cancer

“…It has been found that in MCF-7 cells, the roe protein hydrolysate downregulates COX-2 level (Berkoz et al, 2020 ). In MCF-7 and MDA-MB-231 cells, tuna cooking juice hydrolysate by protease XXIII (PA) with > 2.5 kDa ultrafiltration fraction (PAH 2.5 ) (Hung et al, 2014 ), CPe-III derived from chickpea albumin hydrolysate (Xue et al, 2015 ), and AtMP1 and AtMP2 identified from Anabas testudineus antimicrobial peptides (Najm et al, 2021 ) significantly increase the expressions of p53 and Bax, decrease the level of Bcl-2 which could be overwhelmed by Bax (Yin et al, 1997 ), and upregulate caspases levels. HN-1 also activates p53 and induces a p53-dependent increase of Bax/Bcl-2 ratio in xenograft tumors (Qiao et al, 2019 ).…”

“…Molecular docking analysis showed that Arg1, Gln2, Ala6, Ala8, and Gln9 of CPe-III combine the DNA binding domain of p53 protein (Thr102, Leu111, Asn131, Gln144, Asp228, and Asn268) by hydrogen bonds, resulting in the induction of p53 expression (Xue et al, 2015 ). Najm et al found that hydrogen bonds formed between AtMP1/AtMP2 and p53, Bax, Bcl-2, or caspases (Najm et al, 2021 ). Balsamin is a type I ribosome-inactivating protein purified from Momordica balsamina (Kaur et al, 2012 ).…”

ApInAPDB: a database of apoptosis-inducing anticancer peptides

“…The BCL-2 family is the main proteins that regulate the intrinsic apoptosis pathway while TNFR, FAS (CD95), and DR3/WSL are involved in extrinsic apoptosis 11 . Peptide-based cancer therapy can target both apoptosis pathways 12 , 13 . However, the origin of apoptosis-inducing anticancer peptides is not confined to analogues of the BCL-2 family.…”